Banxia Baizhu Tianma Decoction (BBTD), a classical compound of traditional Chinese medicine (TCM), has been clinically used for hypertension treatment. There are so many components in this formula that it is necessary to explore whether each component plays a positiverole in anti-hypertension treatment. After hypertension models were established with high-fat diet for 6 weeks, rats in each group were treated with full prescription, no principle prescription, no ministerial prescription and no adjuvant prescription for 6 weeks respectively. It was found that BBTD was effective in reducing body weight gain, decreasing serum TG and T-CHO, as well as lowering diastolic blood pressure (DBP) and systolic blood pressure (SBP). Total 28 metabolites in model rats were identified to be different from that of control group, which focusing on 12 important metabolic pathways: (a) Phenylalanine, Tyrosine and tryptophan biosynthesis, (b) Linoleic acid metabolism, (c) Starch and sucrose metabolism, etc. Eight different metabolites (Talose, Oleic acid, L-Proline, etc) were identified between the full prescription group and the model group, which are mainly involved in 4 metabolic pathways (Arginine and proline metabolism, Biosynthesis of unsaturated fatty acids, Amino sugar and nucleotide sugar metabolism and Aminoacyl-tRNA biosynthesis). Furthermore, it also indicated that the principle drugs ie. Pinellia ternata and Gastrodia elata was better than the other drug treatment. In a word, BBTD has a therapeutic effect on hypertension in rats, and Pinellia ternata and Gastrodia elata play an important role in the treatment. The study provides theoretical basis for further optimization of BBTD formula.

Understanding the root traits of tree species is of great importance for selection of trees in dry zone agroforestry systems. We investigated root growth of nine dryland tree species in West Africa and assessed the effect of drought stress on biomass allocation to root systems as function of soil depth. Seedlings were planted in 2m tubes inserted into the soil and subjected to three watering treatments: top irrigation, low irrigation at 50 cm depth, and drought. After 8 and 12 months, we assessed root growth and architecture, and supplemented with assessments of water potential, leaf phenology, and above-ground growth. Results showed large differences in rooting profile of the tree species. Drought induced faster root growth and all species reached the maximum depth after 12 months. Rooting depth was reduced under top irrigation for Adansonia digitata, Sterculia setigera and Anacardium occidentale that concentrated root biomass in the upper soil layers, while Acacia senegal, Acacia nilotica and Faidherbia albida had deeper root systems. Acacia species showed clear differences in leaf water potential between treatments while A. digitata and S. setigera appeared to be water conserving under drought. We discuss results in terms of isohydry/an-isohydry and evergreen vs. deciduous habits.

Background Vertical transmission of SARS-CoV-2 has been reported but appears uncommon. Objectives This study systematically reviewed the evidence on vertical transmission of SARS-CoV-2 from pregnant women to their neonates. Search strategy Literature searches in WHO Covid-19 Database, LitCovid, medRxiv, and Google Scholar for SARS-CoV-2 using keywords and associated synonyms, search date to 20 December 2020; no language restrictions. Selection criteria Studies of any design reporting transmission. Data collection and analysis Two reviewers independently assessed article eligibility and extracted data. Results were reported descriptively; no meta-analyses were possible. Main results 106 studies were included: 40 reviews and 66 primary studies, most conducted in hospitals. 32 case reports were assessed as high risk of bias, due to the study design; across the 34 remaining primary studies, risk of bias was low to moderate. Sixteen case reports described vertical transmission. In cohort studies and case series, 65/2391 (2.7%) neonates born to mothers with a COVID-19 diagnosis tested positive for SARS-CoV-2 within 24 hours of birth; the proportion of positive neonates ranged from 0% to 22%. Twenty studies reported no vertical transmission. Maternal symptomatology and mode of delivery were not correlated with vertical transmission. 7/25 studies of placental tissue identified SARS-CoV-2; vertical transmission was infrequent. No study reported the results of viral culture to detect SARS-CoV-2. Conclusions These findings indicate that vertical transmission is possible, but not frequent. Further high-quality studies are needed to understand vertical transmission. Funding World Health Organization: WHO registration No 2020/1077093.

Objective: To compare the interpregnancy weight gain in women whose first infant had a major congenital anomaly vs. those without an affected child. Design: Nationwide cohort study Setting: Denmark Population: All primigravid women with two consecutive singleton births between January 2004 and December 2017. Main Outcome Measure: Inter-pregnancy weight difference Methods: Multivariable linear regression compared women whose infant had an anomaly vs. those whose infant did not, adjusting for interpregnancy time interval, demographics, smoking and health status at the first pregnancy. Results: Of the 199,536 women, 4035 (2.0%) had a child with an anomaly at the first birth. The mean (SD) maternal BMI at the start of the first pregnancy was 24.2 (4.9) and 23.8 (4.6) kg/m2 in women with, and without, an anomaly-affected newborn. By the start of the second pregnancy, their interpregnancy weight gain was 4.7% and 4.2%, respectively – an adjusted absolute higher gain of 0.34% (95% CI 0.11 to 0.58) or 0.13 kg (95% CI 0.01 to 0.26) in women with an anomaly-affected first-born infant. Compared to those with an unaffected pregnancy, the interpregnancy weight gain difference was greatest in women whose first-born infant had a multi-organ anomaly (0.73%, 95% CI -0.10 to 1.57). Weight gain was more pronounced in women whose infants spent >30 days in hospital between pregnancies. Conclusions: Mothers of an infant with a major congenital anomaly have higher weight gain after pregnancy. Strategies are needed to support a healthy lifestyle in these women.

BACKGROUND: Increased systolic pulmonary artery pressure (sPAP) could lead to mechanical dysfunction and myocardial fibrosis of right heart chambers. Echocardiographic strain analysis has not been adequately studied in patients living with pulmonary hypertension (PH). METHODS AND RESULTS: A cross-sectional cohort of patients with suspected PH and echocardiographic strain evaluation was recruited. Cut-off values of peak tricuspid regurgitation velocity with low probability of PH (≤ 2.8 m/s), intermediate probability (2.9-3.4 m/s, without other echo PH signs) and high probability of PH (2.9-3.4 m/s with other echo PH signs and >3.4 m/s) categories were studied by right ventricular and right atrial strain analysis in a sample of 236 patients, 58 (56.9%) had low, 15 (14.7%) intermediate, and 29 (28.4%) high probability of PH. We observed a negative association between right ventricular free wall strain and atrial global strain with sPAP. As PH severity increased, right atrial reservoir, conduit, and contraction (booster) strain values decreased. Identified cut-off values of strain parameters had an adequate ability to detect PH severity categories In addition, post-mortem biopsies of right heart chambers from subjects with known severe pulmonary hypertension were analyzed to quantify myocardial fibrosis. Our sample of right heart biopsies (n=12) demonstrated an association between increased sPAP before death and right ventricular and right atrial fibrosis. CONCLUSIONS: Mechanical dysfunction and fibrosis in right chambers is associated with increased sPAP. Right ventricular and atrial strain could provide enhancement in the diagnosis and categorization of subjects with suspected PH.

Topological descriptors defined on chemical structures are effective in understanding the properties and activities of chemical molecules. Open neighborhood degree sum based topological indices were studied recently by Vignesh and Kalyani Desikan [29]. Vignesh et al. [30] proposed the reciprocal versions of the indices mentioned in [29]. In this paper, the Quantitative Structure - Property Relationship (QSPR) analysis of the proposed closed neighborhood versions of the indices dened in [29] and [30] are analyzed. QSPR analysis is a statistical procedure for examining molecular activity. QSPR analysis of the proposed indices is performed for octane isomers to uncover their predicting power. Curvilinear regression models are obtained and analysed for the physico-chemical properties of the octane isomers in terms of the proposed indices. We fit linear, quadratic and cubic models for estimating the physico-chemical properties of octane isomers based on the proposed indices. Among our proposed models, the cubic regression model gave the best estimates for the physico-chemical properties of octane isomers. Also, the sensitivity analysis of our proposed indices is performed. Further, we compute the dened closed neighborhood topological indices forHyaluronic Acid - Curcumin Conjugates (HA)n, n>=1.

The design and engineering of promoter architectures with tunable and improved promoter activity is a major challenge that substantially impacts recombinant protein production and industrial biotechnology. Pichia pastoris and Saccharomyces cerevisiae are of industrial interest owing to their recombinant protein production capacities, genetic accessibility, and simple nutrient requirements. We annotated the binding sites for the targeted transcription factors (TFs) in P. pastoris through the constructed yeast curation pipeline using phylogenetic footprints of TF binding sites (TFBSs) in the model yeast S. cerevisiae. The upstream regulatory regions of the genes encoding the enzymes in central carbon metabolism for putative TFBSs in S. cerevisiae and P. pastoris were analyzed with a direct scanning algorithm. The conserved putative TFBSs located in the promoters of the orthologous genes in P. pastoris were identified by the pairwise sequence alignments between the promoters of the two species. Among the putative TFBSs annotated in P. pastoris, the probable functional regions of those in coordination with the TFs play a role in the regulation of central carbon metabolism was evaluated based on the functional regions of their counterparts in S. cerevisiae. The consensus sequence of TFBS motifs in P. pastoris was also determined. The structural analysis we presented is invaluable for designing novel engineered promoters and ensuring transcriptional robustness in P. pastoris and phylogenetic footprinting analyses of other yeast species.

Objectives: Quantify the effect of a non-opioid pain regimen on postoperative opioid prescriptions filled by the patient after adult tonsillectomy. Design: Retrospective cohort study including a time frame before and after a practice change from an opioid/acetaminophen-based postoperative pain regimen to a regimen based on ibuprofen and acetaminophen. Half of study subjects received a prescription for postoperative opioid medications. Half were prescribed ibuprofen following surgery and only provided with opioid analgesia as a rescue medication, where ibuprofen was medically contraindicated, or at patient request following counseling regarding risks of opioids. The New Mexico Prescription Monitoring System was used to verify filled opioid prescriptions. Descriptive statistics and logistic regression were used for data analysis. Settings: Tertiary care academic medical center. Participants: All elective adult tonsillectomies performed consecutively by a single surgeon. Main Outcome Measures: Percent of patients filling an opioid prescription. Results: Ninety-nine patients were included in analysis. 53 received an opioid-based postoperative regimen and 46 received an ibuprofen/acetaminophen-based regimen. There was no difference in the bleeding rate between the two groups. Significantly fewer patients in the ibuprofen group filled postoperative opioid prescriptions when compared to the group that did not receive ibuprofen (40% vs. 96.2%, p<0.0001, OR=0.02). Conclusion: Ibuprofen is a safe and effective analgesic following adult tonsillectomy and significantly reduces the proportion of patients who fill a postoperative opioid prescription.

Epigenetic was identified to closely related to nasal polyp formation in CRSwNP before 18 years ago,epigenetic modifications have gained more attention from researchers owing to their importance role in the pathogenesis of CRSwNP and potential biomarkers for diagnosis and treatment. As regulators, epigenetic modifications (including ncRNA, histone modification and DNA methylation) participate in inflammation and remodeling processes by different signaling pathways through affecting gene expression. In this review, we conclude epigenetic changes in CRSwNP , between ECRSwNP and non-ECRSwNP and CRSwNP with comorbid asthma,and summarize relative signaling pathways.In addition, we review epigenetic changes in response to micro biome factors containing bacterias, virus and environment factors and offer novel epigenetic biomarkers for CRSwNP. Moreover, correlation between clinical presentations and epigenetic changes also is presented. In conclusion, we highlight the importance of studies concerning epigenetic modifications in CRSwNP to help us better understand molecular mechanisms during the development of CRSwNP, and propose some unmeet questions and hypothesis .

We consider a system of non-linear eikonal equations in one space dimension that describes the evolution of interfaces moving with non-signed strongly coupled velocities. We have recently proven the global existence and uniqueness of viscosity solutions for this system, under a BV estimate. In this paper, we propose a semi-explicit scheme that satisfies the same BV estimate proven in the continuous case, at the discrete level, and we show that a certain linear interpolation of the discrete solution to the scheme converges to a viscosity solution of the main system considered. We also provide some numerical simulations in the case of dislocations dynamics

Hydroxyl acid has become an important chemical in the field of materials and medicine due to its dual functional modules. Fortunately, Gluconobacter oxydans whole-cell catalysis is on spotlight with promising potential in bio-catalyzing polyhydroxy chemical to produce hydroxyl acids. Therefore, straight-chain primary diols (C2-C6) were investigated as substrates oxidized by G. oxydans. As results, we found a fantastic critical point of methylene-number determining end-products. G. oxydans catalyzes C4 and smaller methylene-number compounds only forming hydroxyl acids, but C5/C6 can be converted to diacids. Furthermore, it was important that we successfully selective and directionality controlled the product of C5/C6 primary diols to hydroxyl acids instead of diacids through the regulation of pH≥5.5. Finally, we successfully produced nearly 102.3 g/L 5‑hydroxyvaleric acid during 48 h with 99.8% yield by sealed-oxygen supply (SOS) biotechnology which is the highest level. These findings have important reference significance for the selective and directionality bioconversion of primary diols into hydroxyl acids and provide a promising path for the industrial development of hydroxyl acids with integrating C2-C6.

Intraspecific variability (IV) has been proposed as a new track to explain species coexistence. Previous studies generally assumed that IV results from intrinsic differences between conspecifics that widen species’ fundamental niches and blur differences among species, thus impeding stable coexistence, but also slowing down the rate of competitive exclusion. Based on a body of evidence, we here argue that IV does not necessarily imply differences among conspecifics, nor species niches overlap: conspecifics differ in their measured attributes mainly due to differences in the micro-environment they thrive in. Consequently, they respond more similarly to environmental variation than heterospecifics, thereby concentrating competition within species – a necessary condition for species coexistence. We call for new studies exploring observed IV as an outcome of species-specific responses to high-dimensional environmental variations that can lead to inversions of species hierarchy in space and time promoting stable coexistence.

Introduction Ventilation strategies in atrial fibrillation ablation affect procedure outcomes by influencing catheter stability. Studies have highlighted favorable atrial fibrillation (AF) ablation outcomes with the use of high frequency jet ventilation (HFJV) which has shown to improve lesion durability, energy delivery and tissue contact. However, this mode of ventilation is not readily available. In this systematic review we highlight the available data on the use of very low tidal volume, high frequency ventilation using standard ventilators, that aims to provide settings similar to HFJV during AF ablations. Methods Using a combination of search terms in databases and manual searches in bibliographies of identified articles, we reviewed all published data reported in the English language on the use of very low tidal volume with high frequency ventilation during atrial fibrillation ablation. Results A total of 4 manuscripts were identified; 3 cohort studies and 1 case report. The utilization of standard ventilators with a high frequency, very low tidal volume ventilation strategy appears to closely mimic the catheter stability benefits that HFJV ventilators provide. Across the 3 cohort identified studies, the use of this ventilation strategy was associated with improved catheter stability, tissue contact, and decrease in radiofrequency time. No increased risk was identified compared to standard ventilation. Conclusion With a purpose of limiting thoracic excursion and cardiac movement, limited and sparse studies have shown improved outcomes with a very low tidal volume, high frequency ventilation strategy. Additional studies are needed to solidify this easily accessible and widely available mode of ventilation.

A discharge summary is a permanent record of a patient’s hospital visit and the primary means of handover between care providers, but they often lack precision and omit important information. The aim of this study was to develop the ‘ideal’ discharge summary for patients in a regional neurosurgical centre. The essential elements were identified by discussion with clinicians within the department of neurosurgery and summaries were retrospectively audited against these. We then put in place two interventions: a visual aid outlining key components was placed above computer stations in the junior doctors’ offices and formal departmental teaching sessions delivered. After three months we re-audited the discharge summaries retrospectively. Initial audit identified poor documentation especially of medical issues during hospital admission, clinical condition on discharge and safety-netting. After the interventions, documentation in all key components improved dramatically. Good documentation is one of the most important non-clinical aspects of medical practice yet, it is not always part of under-graduate and post-graduate education. Existing guidelines about writing good discharge summaries exist but these are vague and not specific to specialist practice. The development of a simple specialty specific discharge summary guide can improve discharge summary quality and should be encouraged in all specialties.

Introduction. The role of a border zone in arrhythmogenesis is not fully understood. In this study we evaluated independent contributions of action potential duration (APD) and dispersion of repolarization (DOR) across the normal/ischemic border to the development of ventricular tachycardia and/or fibrillation (VT/VF). Methods. Ischemia-reperfusion episodes were induced in anesthetized rats by transient coronary occlusion. Unipolar electrograms were recorded from ischemic and perfused areas using a 64-lead array to obtain activation times (ATs), repolarization times (RTs), activation-repolarization intervals (ARIs, a surrogate for APD) and dispersion of repolarization (DOR, as a difference between the earliest and latest RTs). Pinacidil (0.3 mg/kg) and glibenclamide (2 mg/kg) were applied to reduce DOR and to clamp APD at a lower and upper levels, respectively. Results. In the control animals, APD shortened in the ischemic zone, DOR increased to 9±3 ms, and VT/VF developed at reperfusion (6 out of 10). Pre-occlusion application of glibenclamide prolonged APD in the ischemic and perfused zones, decreased DOR to 5±2 ms and did not affect VT/VF development (4 out of 11). Post-occlusion infusion of pinacidil shortened APD in the perfused zone, decreased DOR to 6±3 ms and VT/VF incidence (2 out of 11). Extrasystolic burden at reperfusion was associated with VT/VF incidence in logistic regression analysis (β=1.182, 95%CI 1.008-1.386, p=0.04) and was lesser (p<0.01) in the pinacidil group as compared to the control and glibenclamide groups. Conclusion. The APDs in the perfused zone were a superior arrhythmogenic factor in respect to DOR in the present ischemia-reperfusion model.

A new shotcrete material with high compressive strength, low water absorption rate and controlled coagulation time was developed. The changes of coagulation time, compressive strength and water absorption rate of shotcrete material with different additions of fly ash were studied according to the traditional proportion of shotcrete materials. The improving effects of the synthetic properties of fly ash shotcrete materials with different maintenance periods were studied by normal temperature activation of the new shotcrete materials through the compounding of alkali activator Na2SO4 and polycarboxylic acid water reducing agent. The results show that it is feasible to use the activated fly ash to make shotcrete material for roadway. The combination of AA-Na2SO4 and PAWRa can improve the comprehensive properties of fly ash shotcrete. When 1.5% and 0.5% of AA-Na2SO4 and PAWRa were added respectively, the comprehensive properties of the shotcrete material were the best.

Aqueous zinc-based flow batteries have received considerable attention for large-scale energy storage due to their low cost, high safety and readily available raw materials. However, zinc dendrite shortens the cycle life, and limited zinc deposits at anodes reduce their capacity. Herein, we design a semi-solid reactor with 3D interconnected electron-ion networks to achieve high areal capacity and uniform distribution of reactive zinc. Multi-layered graphitic felt builds abundant avenues for electron transfer, and zinc slurry offers massive ions transfer channels. The kinetic resistance is 21.5 Ω cm-2, about one-twentieth of that in flood zincate solutions. The capacity is up to 100 mAh cm-2, which is among the highest values in zinc-based flow batteries. The assembled zinc-iron flow battery delivers high coulomb efficiency of 100% and energy efficiency of 85%. Additionally, zinc-iron flow batteries show outstanding cycle stability over 580 hours without performance degradation.

Copper oxide and nickel oxide photocatalysts were synthesized using Abelmoschus esculentus leaf extract and their photocatalytic properties were evaluated by degrading recalcitrant pesticide imidacloprid. A rapid, economical, and environment friendly biosynthesis produced high-surface-area CuO and NiO photocatalyst nanoparticles. The CuO and NiO nanoparticles were characterized by scanning electron microscopy, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and thermo-gravimetric/differential thermal analysis. The XRD studies showed that the synthesized nanoparticles have a face-centered cubic structure. Photocatalytic degradation study showed nanoparticles bear good potential to degrad Imidacloprid. The degradation was found to be affected by the Imidacloprid concentration, solution pH, and photocatalyst dosages. The pesticide degradation percentage (~99%) was attained by CuO photocatalyst that was higher than for NiO photocatalyst (~81%). The degradation follows pseudo-first-order kinetic model, and the apparent rate constant (Kapp) decreased from 0.028/min for CuO to 0.0076/min for NiO photocatalyst. The nanoparticles demonstrated high water stability and recyclability.

The low energy release efficiency seriously hinders the application of Boron (B) powder as metal fuel in solid propellants. To overcome this issue, a series of B-based micro-nanospheres were produced by encapsulating B-based metastable intermixed composites (MICs) with ammonium perchlorate (AP) through a solvent evaporation-induced self-assembly (EISA) method in this work. A self-developed combustion system that contains ignition pool, indicator light, and high-speed camera to precisely monitor ignition delay time and burning time was utilized. The EISA-prepared B/Bi2O3/AP composite show higher combustion temperature (1435 oC) and shorter ignition delay time (379±25 ms) than physically mixed sample (1264 oC and 1035±63 ms). The isolated reaction units of B/Bi2O3/AP-EISA micro-nano composite accelerate heat transfer and accumulation and reduce sintering and agglomeration, thus promoting the combustion efficiency of B powder. This study presents a promising strategy by doping and encapsulating process to increase the combustion performance of B based metal fuel.

The entrainment and accumulation of ice floes in front of the sluice gates are closely related to the water transport efficiency and safe operation of the channel during an ice period. A flume study is carried out for a sluice gate with free outflow. A framework of stacking ensemble models is used to analyze the data, which consists of a two-level structure including the principal component analysis (PCA) and the support vector machine (SVM) algorithms. Based on the mechanism of ice floe accumulation, ten input characteristics of the machine learning (ML) model are selected. The PCA method is used to eliminate redundant information. The first principal component, with a contribution rate of 71.76%, and the second principal component, with a contribution of rate 15.64%, are extracted as the inputs of the SVM model, and the state of the floating ice in front of the gate is used to determine the classification labels. The 5-fold cross-validation method is used to train the model. The training results showed that the Gaussian radial basis functions (RBF) were the optimal kernel function. The performance of the developed model is measured using area under curve (AUC), accuracy (Acc) and F1-score (F1) values as statistical indicators. The results showed that the established PCA-SVM model improves the Bernoulli naive Bayes (Bernoulli NB) classifier and K-nearest neighbors’ algorithm (KNN) models. It increasing the AUC value by 11% and 5%, the Acc value by 16% and 17%, and the F1 value by 17% and 2%, respectively.

Objective To identify when obstetricians would deliver a fetus with antenatal hydronephrosis and normal liquor Design Snap-shot survey Setting Survey Monkey link Population or Sample Obstetrics and fetal medicine consultants who received the survey link via closed professional forums on the North West Coast Maternity Clinical Network, Facebook, and publicly on Twitter Methods Survey link publicised as above, obstetric consultants were asked at what gestation would they deliver a fetus with antenatal hydronephrosis and normal liquor; and what criteria would they use to make that decision Main Outcome Measures Number of years in practice, gestation at delivery, anteroposterior diameter (APD) of renal pelvis Results 44/102 respondents (43%) would deliver prior to 40 weeks (median no of years as consultant 10 years (IQR 5-17) versus those who would not median years as consultant 5.5 (3-12). Re APD threshold of delivery: 17 indicated delivery if the APD were 20mm, 10if it were 21-30mm and 16 if it were >30mm. Re gestation at which they would deliver: 13 indicated 37-38weeks, 13 indicated 38-39 weeks and 17 indicated 39-40 weeks. Reasons selected for delivery before term were obstetric anxiety n=2, maternal request n=2, maternal anxiety n=2 and concern about fatal renal damage / renal n=34. Conclusions A surprising number of respondents would consider early delivery of a fetus with hydronephrosis and normal liquor despite the lack of evidence of benefit. The evidence supporting term delivery means that early term delivery is only indicated for obstetric reasons in this scenario.

Mendelian susceptibility to mycobacterial disease (MSMD) is a group of genetic disorders characterized by a defect in interferon γ (IFN γ)–mediated immunity, with predisposition to infections caused by atypical and low virulent mycobacteria. We describe a 10-year-old boy, presented with left inguinal and submandibular lymphadenopathy with low grade fever for 10 days. Past history was significant and at the age of 4 months and 5 years, he was diagnosed to have tubercular axillary lymphadenitis and was treated with anti-tubercular drugs on each occasion. On examination, he had left submandibular and inguinal lymphadenopathy and mild splenomegaly. He underwent left inguinal lymph node biopsy that reported granulomatous lymphadenitis and culture reported growth of Salmonella enterica. Genetic evaluation showed pathogenic homozygous mutation of IL12Rß1 gene. A diagnosis of MSMD was established and he was treated with antibiotics to which he responded promptly. A detailed review of literature on IL12RB1 deficiency was performed. Index case has been discussed in light of the previously published literature.

Newborn screening is an important public health measure of secondary prevention. With the increasing number of target conditions, there is a growing demand to optimize laboratory processes to save patient material and to work cost-effectively. Here, we report an adaption of the commercially available SpotOn Clinical Diagnostics tandem mass spectrometry test kit to detect hemoglobin fragments as well as substrate-product pairs of biotinidase and porphobilinogen synthase at once. The presence of specific peptides and the enzyme activities allows to infer to disease states, i.e., sickle cell disease, biotinidase deficiency and hereditary tyrosinemia type 1.

For the production of recombinant protein therapeutics in mammalian cells, a high rate of gene expression is desired and hence strong viral-derived promoters are commonly used. However, they usually induce cellular stress and can be susceptible to epigenetic silencing. Endogenous promoters, which coordinates their activity with cellular and bioprocess dynamics while at the same time they maintain high expression levels, may help to avoid such drawbacks. In this work, endogenous promoters were identified based on high expression levels in RNA-seq data of CHO-K1 cells cultured in high density. The promoters of Actb, Ctsz, Hmox1, Hspa5, Vim and Rps18 genes were selected for generating new expression vectors for the production of recombinant proteins in mammalian cells. The in silico-derived promoter regions were experimentally verified and the majority showed transcriptional activity comparable or higher than CMV. Also, stable expression following a reduction of culture temperature was investigated. The characterized endogenous promoters (excluding Rps18) constitute a promising alternative to CMV promoter due to their high strength, long-term expression stability and integration into the regulatory network of the host cell. These promoters may also comprise an initial panel for designing cell engineering strategies and synthetic promoters, as well as for industrial cell line development.